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AB257580

Human PFKP knockout HeLa cell lysate

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PFKP KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon3.
2 Images
Western blot - Human PFKP knockout HeLa cell lysate (AB257580)
  • WB

Lab

Western blot - Human PFKP knockout HeLa cell lysate (AB257580)

Lane 1 : Wild-type HeLa cell lysate (20 ug)

Lane 2 : PFKP knockout HeLa cell lysate (20 ug)

Lane 3 : Jurkat cell lysate (20 ug)

Lanes 1-3 : Merged signal (red and green). Green - ab119796 observed at 86 kDa. Red - loading control ab181602 observed at 36 kDa.

ab119796 Anti-PFKP antibody [OTI1D6] was shown to specifically react with PFKP in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265136 (knockout cell lysate ab257580) was used. Wild-type and PFKP knockout samples were subjected to SDS-PAGE. ab119796 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-PFKP antibody [OTI1D6] (<a href='/en-us/products/primary-antibodies/pfkp-antibody-oti1d6-ab119796'>ab119796</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PFKP knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PFKP knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-pfkp-knockout-hela-cell-line-ab265136'>ab265136</a>)

Lane 3:

Jurkat cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution

Predicted band size: 86 kDa

Observed band size: 86 kDa

false

Sanger Sequencing - Human PFKP knockout HeLa cell lysate (AB257580)
  • Sanger seq

Unknown

Sanger Sequencing - Human PFKP knockout HeLa cell lysate (AB257580)

Homozygous : 1 bp insertion in exon3

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon3.

Disease

Adenocarcinoma

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Complementary Products

Primary Antibodies

AB204131

Anti-PFKM + PFKP antibody [EPR17314]

RabMAb|Recombinant

Flow Cytometry (Intracellular) - Anti-PFKM + PFKP antibody [EPR17314] (AB204131)

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Assay Kits

AB155898

Phosphofructokinase Activity Assay Kit (6-PFK, Colorimetric)

Functional Studies - Phosphofructokinase Activity Assay Kit (6-PFK, Colorimetric) (AB155898)

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Proteins & Peptides

AB259412

Recombinant human VEGF 165A protein (Active)

Premium Bioactive

Mass Spectrometry - Recombinant human VEGF 165A protein (Active) (AB259412)

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Proteins & Peptides

AB180317

Recombinant Human POLR1C protein

SDS-PAGE - Recombinant Human POLR1C protein (AB180317)

  • 0
  • 0
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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

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What's included?

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Properties and storage information

Gene name
PFKP
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

PFKP also known as 6-phosphofructokinase platelet type functions as an important glycolytic enzyme that catalyzes the phosphorylation of fructose 6-phosphate to fructose 16-bisphosphate using ATP. This process shows the rate-limiting step in glycolysis. The PFKP protein weighs approximately 85 kDa. Expression occurs in various tissues with high presence in platelets and muscle tissues indicating its significant role in energy metabolism.
Biological function summary

PFKP plays an essential role in maintaining energy supply ensuring efficient glycolysis during high energy demands. It associates with other isoforms PFKM (muscle) and PFKL (liver) to form a tetrameric complex which can adapt its activity based on the energy needs of the cell. This adaptability defines its contribution to cellular metabolism regulation considering energy substrates and signaling molecules.

Pathways

The function of PFKP ties closely to glycolysis and the larger metabolic pathways managing energy production such as the tricarboxylic acid (TCA) cycle. Within these pathways PFKP interacts with other metabolic regulators including AMP-activated protein kinase (AMPK) which influences cell metabolism in response to energy availability. Connections with PFKL and PFKM reveal that PFKP's activity relates to the overall glycolytic flux and energy balance.

PFKP's alterations contribute to metabolic conditions like cancer and diabetes. Its dysregulation in cancer cells supports the Warburg effect where tumor cells prefer glycolysis. The link with metabolic proteins such as AMPK also indicates a potential role in diabetes as both disorders involve shifts in energy metabolism. Understanding PFKP's impact on these diseases helps in developing therapeutic strategies targeting metabolic pathways.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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